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Transient Fluorescence Labeling: Low Affinity—High Benefits
Fluorescent labeling is an established method for visualizing cellular structures and dynamics. The fundamental diffraction limit in image resolution was recently bypassed with the development of super-resolution microscopy. Notably, both localization microscopy and stimulated emission depletion (ST...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8583718/ https://www.ncbi.nlm.nih.gov/pubmed/34769228 http://dx.doi.org/10.3390/ijms222111799 |
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author | Perfilov, Maxim M. Gavrikov, Alexey S. Lukyanov, Konstantin A. Mishin, Alexander S. |
author_facet | Perfilov, Maxim M. Gavrikov, Alexey S. Lukyanov, Konstantin A. Mishin, Alexander S. |
author_sort | Perfilov, Maxim M. |
collection | PubMed |
description | Fluorescent labeling is an established method for visualizing cellular structures and dynamics. The fundamental diffraction limit in image resolution was recently bypassed with the development of super-resolution microscopy. Notably, both localization microscopy and stimulated emission depletion (STED) microscopy impose tight restrictions on the physico-chemical properties of labels. One of them—the requirement for high photostability—can be satisfied by transiently interacting labels: a constant supply of transient labels from a medium replenishes the loss in the signal caused by photobleaching. Moreover, exchangeable tags are less likely to hinder the intrinsic dynamics and cellular functions of labeled molecules. Low-affinity labels may be used both for fixed and living cells in a range of nanoscopy modalities. Nevertheless, the design of optimal labeling and imaging protocols with these novel tags remains tricky. In this review, we highlight the pros and cons of a wide variety of transiently interacting labels. We further discuss the state of the art and future perspectives of low-affinity labeling methods. |
format | Online Article Text |
id | pubmed-8583718 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-85837182021-11-12 Transient Fluorescence Labeling: Low Affinity—High Benefits Perfilov, Maxim M. Gavrikov, Alexey S. Lukyanov, Konstantin A. Mishin, Alexander S. Int J Mol Sci Review Fluorescent labeling is an established method for visualizing cellular structures and dynamics. The fundamental diffraction limit in image resolution was recently bypassed with the development of super-resolution microscopy. Notably, both localization microscopy and stimulated emission depletion (STED) microscopy impose tight restrictions on the physico-chemical properties of labels. One of them—the requirement for high photostability—can be satisfied by transiently interacting labels: a constant supply of transient labels from a medium replenishes the loss in the signal caused by photobleaching. Moreover, exchangeable tags are less likely to hinder the intrinsic dynamics and cellular functions of labeled molecules. Low-affinity labels may be used both for fixed and living cells in a range of nanoscopy modalities. Nevertheless, the design of optimal labeling and imaging protocols with these novel tags remains tricky. In this review, we highlight the pros and cons of a wide variety of transiently interacting labels. We further discuss the state of the art and future perspectives of low-affinity labeling methods. MDPI 2021-10-30 /pmc/articles/PMC8583718/ /pubmed/34769228 http://dx.doi.org/10.3390/ijms222111799 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Review Perfilov, Maxim M. Gavrikov, Alexey S. Lukyanov, Konstantin A. Mishin, Alexander S. Transient Fluorescence Labeling: Low Affinity—High Benefits |
title | Transient Fluorescence Labeling: Low Affinity—High Benefits |
title_full | Transient Fluorescence Labeling: Low Affinity—High Benefits |
title_fullStr | Transient Fluorescence Labeling: Low Affinity—High Benefits |
title_full_unstemmed | Transient Fluorescence Labeling: Low Affinity—High Benefits |
title_short | Transient Fluorescence Labeling: Low Affinity—High Benefits |
title_sort | transient fluorescence labeling: low affinity—high benefits |
topic | Review |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8583718/ https://www.ncbi.nlm.nih.gov/pubmed/34769228 http://dx.doi.org/10.3390/ijms222111799 |
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